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Traditional methods for mining novel biological enzymes are mainly based on functional screening of culturable microorganisms followed by sequence analysis and functional validation of positive strains. Enzymes such as amylases and esterases were discovered using traditional mining methods. Microorganisms play a central role in traditional methods as the main source of catalysts. However, most microorganisms exist in natural microbial community environments, are interdependent with other microorganisms, and are difficult to culture in pure form under laboratory conditions, which greatly limits the efficiency of traditional methods for mining novel enzymes. Sequence-based enzyme screening technology has emerged, which can accurately and efficiently screen enzyme sequences with specific functions from large-scale biological sample libraries by utilizing high-throughput sequencing technology and computational algorithms. This technology breaks through the limitations of traditional screening methods and opens up new possibilities for the optimization of enzyme catalytic systems and the development of new enzymes.
In today's rapidly developing biotechnology, enzymes play an important role in medicine, agriculture, energy, and industry. Through enzyme screening, we can efficiently identify and optimize enzymes with specific functions and properties. Creative Enzymes specializes in innovative enzyme screening methods, and sequence-based enzyme screening services are one of our core businesses.
Creative Enzymes' sequence-based enzyme screening services are designed to meet the precise needs of our clients for innovative enzymes. Our service process is as follows:
1. Project design
Our team of experts will work with the client to fully understand the requirements of the project in terms of background, objectives, and intended application. Based on this information, we will develop a targeted enzyme screening program and provide detailed technical consultation to the client.
2. Sample preparation
Customers can provide their own enzyme samples, or we can provide a diverse range of starting samples from our repository upon request. We can also customize the synthesis of sequence-specific enzyme samples if required.
3. Designing enzyme variant libraries
We design and construct libraries of enzyme variants according to our clients' needs and goals. This can be directed mutation (point mutation, insertion, deletion, etc.) or random mutation using a library of known enzyme genes.
5. High-throughput sequencing
We sequence enzyme variant libraries using advanced high-throughput sequencing technologies (e.g. Illumina HiSeq or MiSeq) to obtain large amounts of sequence data.
6. Sequence alignment and analysis
Using bioinformatics tools and algorithms, we perform sequence comparison and analysis (including variant analysis, structure prediction, function prediction, etc.) of the sequencing data to determine the nature and potential functions of the enzyme variants.
7. Screening and evaluation
We will select a number of the most promising enzymes from the candidate enzyme sequences and conduct further enzyme activity screening and property assessment, which includes enzyme activity screening, affinity screening, stability screening, etc., in order to select the enzyme variants with excellent performance.
8. Project report
We will document the screening process and results in detail and provide a comprehensive project report to the client. The report will contain enzyme sequence information, activity evaluation results, experimental data and analysis, and recommendations for further applications.
If you are interested in sequencing-based enzyme screening services or have any questions, please feel free to contact us. Our team will be happy to provide you with support and answers.
Figure 1 Pairwise sequence alignment-based strategy for new enzyme discovery
(RSC Catalysis Series 2018)
